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Nucleic acid molecules and other molecules associated with plants

Inactive Publication Date: 2008-05-01
CONNER TIMOTHY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0030] The present invention also provides a method of producing a plant containing one or more proteins encoded by sequences comprising SEQ ID NO: 1 or complement thereof through SEQ ID NO: 18718 or complements thereof, expressed in a sufficient amount and / or fashion to produce a desirable agronomic effect.
[0035] (iii) a 3′ non-translated DNA sequence which functions in plant cells to cause the addition of polyadenylated nucleotides to the 3′ end of RNA sequence; where the promoter is homologous or heterologous with respect to the coding sequence and adapted to cause sufficient expression of a protein in desired plant tissues to enhance the agronomic utility of a plant transformed with said gene.

Problems solved by technology

Automated single run sequencing typically results in an approximately 2-3% error or base ambiguity rate.
BLOSUM62 is tailored for alignments of moderately diverged sequences and thus may not yield the best results under all conditions.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0241] The cDNA library of the present invention designated LIB9, is prepared from Arabidopsis thaliana, Columbia ecotype, leaf tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. Leaf blades are cut with sharp scissors at seven weeks after planting. The tissue is immediately frozen in liquid nitrogen and stored at −80° C. until total RNA extraction. SEQ ID NO: 1 to SEQ ID NO: 1425 are from LIB9.

[0242] The cDNA library of the present invention designated LIB22, is prepared from Arabidopsis thaliana Columbia ecotype root tissue. Wild type Arabidopsis thaliana seeds are planted in commonly used planting pots and grown in an environmental chamber. After 5-6 weeks the plants are in the reproductive growth phase. Stems are bolting from the base of the plants. After 7 weeks, more stems and floral buds appear, and a few flowers are starting to open. Roots of 7-week old plants from pots are rinsed intensively with t...

example 2

[0248] The stored RNA is purified using Trizol reagent from Life Technologies (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.), essentially as recommended by the manufacturer. Poly A+ RNA (mRNA) is purified using magnetic oligo dT beads essentially as recommended by the manufacturer (Dynabeads, Dynal Corporation, Lake Success, N.Y. U.S.A.).

[0249] Construction of plant cDNA libraries is well-known in the art and a number of cloning strategies exist. A number of cDNA library construction kits are commercially available. The Superscript™ Plasmid System for cDNA synthesis and Plasmid Cloning (Gibco BRL, Life Technologies, Gaithersburg, Md. U.S.A.) is used, following the conditions suggested by the manufacturer.

example 3

[0250] The cDNA libraries are plated on LB agar containing the appropriate antibiotics for selection and incubated at 37° for a sufficient time to allow the growth of individual colonies. Single colonies are individually placed in each well of a 96-well microtiter plates containing LB liquid including the selective antibiotics. The plates are incubated overnight at approximately 37° C. with gentle shaking to promote growth of the cultures. The plasmid DNA is isolated from each clone using Qiaprep plasmid isolation kits, using the conditions recommended by the manufacturer (Qiagen Inc., Santa Clara, Calif. U.S.A.).

[0251] The template plasmid DNA clones are used for subsequent sequencing. For sequencing the cDNA libraries of LIB9, LIB22, LIB23, LIB24, LIB25, LIB35, and LIB146, a commercially available sequencing kit, such as the ABI PRISM dRhodamine Terminator Cycle Sequencing. Ready Reaction Kit with AmpliTaq® DNA Polymerase, FS, is used under the conditions recommended by the manuf...

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PUM

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Abstract

Expressed Sequence Tags (ESTs) isolated from Arabidopsis thaliana are disclosed. The ESTs provide a unique molecular tool for the targeting and isolation of novel genes for plant protection and improvement. The disclosed ESTs have utility in the development of new strategies for understanding critical plant developmental and metabolic pathways. The disclosed ESTs have particular utility in isolating genes and promoters, identifying and mapping the genes involved in developmental and metabolic pathways, and determining gene function. Sequence homology analyses using the ESTs provided in the present invention, will result in more efficient gene screening for desirable agronomic traits. An expanding database of these select pieces of the plant genomics puzzle will quickly expand the knowledge necessary for subsequent functional validation, a key limitation in current plant biotechnology efforts.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is a continuation under 35 U.S.C. § 120 of U.S. application Ser. No. 09 / 333,534 filed Jun. 14, 1999, the entirety of which is herein incorporated by reference.INCORPORATION OF SEQUENCE LISTING [0002] This application contains a sequence listing, which is contained on three identical CD-ROMs: two copies of the sequence listing (Copy 1 and Copy 2) and a sequence listing Computer Readable Form (CRF), all of which are herein incorporated by reference. All three CD-ROMs each contain one file called “15404C seq list.txt” which is 14,348,288 bytes in size (measured in Windows XP) and which was created on Dec. 21, 2005. FIELD OF THE INVENTION [0003] The present invention is in the field of plant biochemistry. More specifically the invention relates to nucleic acid molecules that encode proteins and fragments of proteins produced in plant cells, in particular, Arabidopsis thaliana plants. The invention also relates to proteins an...

Claims

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Application Information

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IPC IPC(8): A01H1/00A01H5/00C07H21/04C12N15/87
CPCC07K14/415
Inventor CONNER, TIMOTHYRUAN, YIJUN
Owner CONNER TIMOTHY
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